Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.

Objective To investigate the regulatory effects and molecular mechanisms of PUS3 on GBM cell malignant behaviors(proliferation,apoptosis,invasion)in vitro,providing potential therapeutic targets for GBM.Methods The expression of PUS3 in GBM was analyzed using the GEPIA2 database.Kaplan-Meier survival analysis evaluated the survival difference between PUS3 high-and low-expression patients.qRT-PCR and Western blot were performed to detect PUS3 expression in normal glial cells(HEB)and GBM cell lines(U87,LN229,U251,T98G).PUS3-stably overexpressing GBM cell lines were constructed.Colony formation,CCK-8 assay,and flow cytometry were used to assess proliferation and apoptosis.Transwell assay evaluated cell invasion.Immunohis-tochemistry(IHC)detected PUS3 expression in GBM patient tissues.Western blot analyzed tumor-related pathway proteins after PUS3 overexpression.Results The expression level of PUS3 is elevated in GBM patient tissues,and the mRNA and protein expression levels in GBM cells are significantly higher than those in HEB cells(P<0.01).After overexpression of PUS3,the proliferation ability of GBM cells was enhanced(P<0.05),the apoptosis rate decreased(P<0.01),and the number of invasive cells increased(P<0.001).Mechanistically,overexpression of PUS3 significantly activates the AKT pathway,and the use of AKT inhibitors in PUS3 overexpressing cells can reverse the pro cancer effect.Conclusion PUS3 is highly expressed in glioblastoma(GBM)and promotes tumor cell proliferation,invasion,and apoptosis inhibition by activating the AKT pathway,suggesting its potential as a therapeutic target for GBM treatment.

Objective To evaluate the effect of two different acid etching agents on the bonding strength between deciduous enamel and composite resin at different etching times.Methods Seventy primary incisors were made into test specimens and randomly divided into 7 groups(n=10),6 experimental groups,and 1 blank control group.The specimens were divided into groups and etchedwith two different acid etching agents(35％H3PO4,15％HCl)for different durations(15,30,60 s).After bonding with the composite resin,shear force testing was performed.Additionally,two deciduous molar dental crowns were cut vertically along the gingival axis,with each tooth divided into seven sections and randomly assigned to seven groups.Enamel acid etched specimens were produced and the surface morphology was observed using scanning electron microscopy.Finally,24 deciduous molars were randomly divided into 3 groups(n=8)according to the etching time(15,30,60 s),and each tooth was vertically cut into 2 pieces along the buccal lingual direction,namely the phosphoric acid group and the hydrochloric acid group,and etched for the same time(n=8),for a total of 6 groups.They were bonded with resin and made into specimens.The specimen was cut vertically to the bonding surface into 2 parts.One measurement point was selected for each part,and each group had a total of 16 measurement values.Resin protrusion length was measured under scanning electron microscopy and was statistically analyzed.Results When comparing acid etching for 15,30,and 60 s,the bonding strength of the phosphate group was higher than that of the hydrochloric acid group.Under scanning electron microscopy,there was no significant difference in the acid etching mode between the two groups,both of which were type 2 acid etching modes dissolved along the glaze column direction.At 15 seconds of acid etching,the enamel surface was uneven,and continuous and uniform dissolution ap-peared at 30 seconds.The length of resin protrusion increased with the increase of acid etching time,and the phosphoric acid group was greater than the hydrochloric acid group at 15,30,and 60 s compared between groups.Conclusion The etching time is positively correlated with the shear bonding strength.The best bonding effect is achieved when deciduous teeth are etched with 35％ H3PO4 for 60 seconds,and the effect of 35％phosphoric acid etching on deciduous tooth enamel is better than that of 15％hydrochloric acid.

Objective This study aims to explore the application effectiveness of the PDCA cycle method in preventing and reducing internal errors in the intravenous drug dispensing center(PIVAS).Methods Internal error data from our hospital's PIVAS in 2020 and 2021 were collected.The data from 2020 represented the pre-implementation of the PDCA cycle,while the data from 2021 represented the post-implementation period.The changes in internal errors and error rates before and after imple-mentation were compared.Results After implementing the PDCA cycle management measures,the annual error rate decreased from 0.887％o in 2020 to 0.681‰ in 2021,a decrease of 23.22％.Conclusion The PDCA cycle method can effectively reduce the occurrence of internal errors,improve work accuracy,and ensure the safety of clinical drug use in PIVAS.

Objective To evaluate the effect of two different acid etching agents on the bonding strength between deciduous enamel and composite resin at different etching times.Methods Seventy primary incisors were made into test specimens and randomly divided into 7 groups(n=10),6 experimental groups,and 1 blank control group.The specimens were divided into groups and etchedwith two different acid etching agents(35％H3PO4,15％HCl)for different durations(15,30,60 s).After bonding with the composite resin,shear force testing was performed.Additionally,two deciduous molar dental crowns were cut vertically along the gingival axis,with each tooth divided into seven sections and randomly assigned to seven groups.Enamel acid etched specimens were produced and the surface morphology was observed using scanning electron microscopy.Finally,24 deciduous molars were randomly divided into 3 groups(n=8)according to the etching time(15,30,60 s),and each tooth was vertically cut into 2 pieces along the buccal lingual direction,namely the phosphoric acid group and the hydrochloric acid group,and etched for the same time(n=8),for a total of 6 groups.They were bonded with resin and made into specimens.The specimen was cut vertically to the bonding surface into 2 parts.One measurement point was selected for each part,and each group had a total of 16 measurement values.Resin protrusion length was measured under scanning electron microscopy and was statistically analyzed.Results When comparing acid etching for 15,30,and 60 s,the bonding strength of the phosphate group was higher than that of the hydrochloric acid group.Under scanning electron microscopy,there was no significant difference in the acid etching mode between the two groups,both of which were type 2 acid etching modes dissolved along the glaze column direction.At 15 seconds of acid etching,the enamel surface was uneven,and continuous and uniform dissolution ap-peared at 30 seconds.The length of resin protrusion increased with the increase of acid etching time,and the phosphoric acid group was greater than the hydrochloric acid group at 15,30,and 60 s compared between groups.Conclusion The etching time is positively correlated with the shear bonding strength.The best bonding effect is achieved when deciduous teeth are etched with 35％ H3PO4 for 60 seconds,and the effect of 35％phosphoric acid etching on deciduous tooth enamel is better than that of 15％hydrochloric acid.

Objective To investigate the regulatory effects and molecular mechanisms of PUS3 on GBM cell malignant behaviors(proliferation,apoptosis,invasion)in vitro,providing potential therapeutic targets for GBM.Methods The expression of PUS3 in GBM was analyzed using the GEPIA2 database.Kaplan-Meier survival analysis evaluated the survival difference between PUS3 high-and low-expression patients.qRT-PCR and Western blot were performed to detect PUS3 expression in normal glial cells(HEB)and GBM cell lines(U87,LN229,U251,T98G).PUS3-stably overexpressing GBM cell lines were constructed.Colony formation,CCK-8 assay,and flow cytometry were used to assess proliferation and apoptosis.Transwell assay evaluated cell invasion.Immunohis-tochemistry(IHC)detected PUS3 expression in GBM patient tissues.Western blot analyzed tumor-related pathway proteins after PUS3 overexpression.Results The expression level of PUS3 is elevated in GBM patient tissues,and the mRNA and protein expression levels in GBM cells are significantly higher than those in HEB cells(P<0.01).After overexpression of PUS3,the proliferation ability of GBM cells was enhanced(P<0.05),the apoptosis rate decreased(P<0.01),and the number of invasive cells increased(P<0.001).Mechanistically,overexpression of PUS3 significantly activates the AKT pathway,and the use of AKT inhibitors in PUS3 overexpressing cells can reverse the pro cancer effect.Conclusion PUS3 is highly expressed in glioblastoma(GBM)and promotes tumor cell proliferation,invasion,and apoptosis inhibition by activating the AKT pathway,suggesting its potential as a therapeutic target for GBM treatment.

Objective This study aims to explore the application effectiveness of the PDCA cycle method in preventing and reducing internal errors in the intravenous drug dispensing center(PIVAS).Methods Internal error data from our hospital's PIVAS in 2020 and 2021 were collected.The data from 2020 represented the pre-implementation of the PDCA cycle,while the data from 2021 represented the post-implementation period.The changes in internal errors and error rates before and after imple-mentation were compared.Results After implementing the PDCA cycle management measures,the annual error rate decreased from 0.887％o in 2020 to 0.681‰ in 2021,a decrease of 23.22％.Conclusion The PDCA cycle method can effectively reduce the occurrence of internal errors,improve work accuracy,and ensure the safety of clinical drug use in PIVAS.

Objective:To evaluate the efficacy and safety of the gemcitabine combined with oxaliplatin (GEMOX) regimen in the postoperative adjuvant treatment for the patients with cisplatin-intolerant uroepithelial cancer.Methods:The clinical data of 98 patients with uroepithelial carcinoma intolerant to cisplatin chemotherapy who underwent radical surgery from August 2017 to October 2022 at Drum Tower Hospital of Nanjing University School of Medicine were retrospectively analysed. The patients were divided into the adjuvant chemotherapy group and the observation group according to whether or not they underwent adjuvant chemotherapy after surgery. The adjuvant chemotherapy group received postoperative chemotherapy with the GEMOX regimen (gemcitabine 1 000 mg/m 2 intravenously on days 1 and 8, oxaliplatin 130 mg/m 2 intravenously on day 2, every 3 weeks as a cycle), and the observation group did not undergo postoperative adjuvant chemotherapy. In the adjuvant chemotherapy group, there were 33 males and 10 females, the patients’ age was (67.8±7.3) years old, 33 cases with estimated glomerular filtration rate (eGFR) ≤60 ml/(min·1.73m 2), and 10 cases with a Eastern Cooperative Oncology Group (ECOG) functional status score of >1. The postoperative pathology showed 39 cases were in stage T 3, 4 cases in stage T 4, and lymph node positivity (N+ ) was found in 10 cases. There were 55 cases in the observation group, with 42 males and 13 females and the age of (70.7±7.7) years old. Forty-two of them had an eGFR ≤60 ml/(min·1.73m 2), and 13 of them had a ECOG score of >1. The postoperative pathology showed 48 cases were in stage T 3, 7 cases in stage T 4, and 13 cases of N+. The changes in renal function, ECOG scores, and adverse reactions were observed in adjuvant chemotherapy group. Kaplan-Meier method was used to estimate the survival rate, and the log-rank test was used to compare the survival rate between groups. Multifactorial Cox regression was used to analyse the correlation between age, lymph nodes, whether or not to combine with adjuvant chemotherapy and the survival of patients. Results:All patients in this study were followed up for 3 to 75 months, with a median follow-up time of 22 (14, 34) months. The recurrence rates were 83.6%(46/55) and 65.1%(28/43) in the observation and adjuvant chemotherapy groups, respectively, and the disease mortality rates were 52.7%(29/55) and 27.9%(12/43), respectively. The results of the Kaplan-Meier survival analyses showed that the 1-, 2- and 3-year disease-free survival rates in the adjuvant chemotherapy group were 62.8%, 48.6% and 41.1%, respectively, and the 1-, 2- and 3-year overall survival rates were 86.0%, 79.0% and 76.4%, respectively. The 1-, 2- and 3-year disease-free survival rates of the observation group were 58.2%, 22.6% and 9.6%, respectively, and the 1-, 2- and 3-year overall survival rates were 78.2%, 49.4% and 42.8%, respectively. The adjuvant chemotherapy group had an advantage over the observation group regarding disease-free and overall survival rates (all P<0.05). The results of multifactorial Cox regression analysis suggested that the functional status score and the presence or absence of positive lymph nodes, diabetes mellitus, and co-adjuvant chemotherapy were independent risk factors affecting the survival of the patients ( P<0.05). Forty-three cases had 1 to 6 courses of adjuvant chemotherapy, with a median course of 4 (2, 4). In terms of safety, the most common adverse reaction in the gastrointestinal tract was loss of appetite (53.4%, 23/43), the most common grade 1 to 2 adverse reaction in myelosuppression was a decrease in haemoglobin (51.2%, 22/43), and the most common grade 3 to 4 adverse reaction was thrombocytopenia (9.3%, 4/43). The eGFR of 33 patients with renal insufficiency in the adjuvant chemotherapy group was higher after each administration cycle than before ( P<0.05), and renal function did not deteriorate with the increase in administration cycles. Ten patients with a ECOG score of 2 remained with a score of 2 after chemotherapy. Conclusions:In patients with cisplatin-intolerant uroepithelial cancer, gemcitabine in combination with an oxaliplatin regimen improves the overall survival of patients. At the same time, it is well tolerated without increasing nephrotoxicity, making it an optional postoperative adjuvant treatment for patients with cisplatin-intolerant uroepithelial cancer.

Objective To investigate the effect of tiopronin on renal function during anti-tuberculosis liver protection therapy.Methods Clinical data of patients with initially treated sensitive tuberculosis treated in our hos-pital from September 2019 to September 2022 and whose anti-tuberculosis regimen was only isoniazid,rifampicin,pyrazinamide and ethambutol were retrospectively analyzed.The patients were divided into study group and control group according to whether tiopronin was used.The baseline data,blood creatinine(Scr),blood urea nitrogen(BUN),urine protein,creatinine clearance,drug combination and related adverse reactions of the two groups were compared.Results Patients obtained based on inclusion and exclusion criteria were divided into a study group(n=102)(antitubercular drugs+tiopronin)and a control group(n=105)(antitubercular drugs+glutathione).There were no statistically significant differences(P＞0.05)in ALT,AST,DBIL,and TBIL levels between the two groups before treatment,at Middle and late treatment.At the later stage of treatment,serum Scr,BUN,creatinine clearance and urinary protein showed statistical differences between the study group and the control group(P＜0.05).The abnormal rate of indicators and the incidence of adverse reactions in the study group were higher than those in the control group at the later stage of treatment(P＜0.05).Conclusion For patients undergoing tuberculosis treatment,the efficacy of tiopronin and glutathione in protecting the liver is comparable.However,in terms of renal function,long-term use of tiopronin is associated with more pronounced damage.Due to the relatively low cost of tiopronin,for families with heavy economic burdens,short-term use of the drug can ensure safety,while long-term use requires close monitoring of renal function changes and timely adjustments to medication.